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PMID: 1330154 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Vasopressin-stimulated [3H]-inositol phosphate and [3H]-phosphatidylbutanol accumulation in A10 vascular smooth muscle cells.

British journal of pharmacology ·Vol. 107 ·No. 1 ·1992-09-00 ·Pages 109-15

Plevin R, Stewart A, Paul A, Wakelam MJ

Abstract

1. The characteristics of vasopressin-stimulated phosphatidylinositol 4,5 bisphosphate (PtdIns(4,5)P2) and phosphatidylcholine (PtdCh) hydrolysis were examined in A10 vascular smooth muscle cells (VSMC), by assessing the formation of [3H]-inositol phosphates ([3H]-IP) and the accumulation of the phospholipase D (PLD) specific product, [3H]-phosphatidylbutanol ([3H]-PtdBuOH). 2. Vasopressin ([Arg8]-VP) and a number of related analogues stimulated the accumulation of [3H]-IP and [3H]-PtdBuOH with similar EC50 values, generating the same rank order of potency for each response (Arg8-VP = vasotocin = Lys8-VP much greater than oxytocin). 3. Inhibition of vasopressin-stimulated [3H]-IP and [3H]-PtdBuOH accumulation by the V1a receptor antagonists, Des-Gly9[beta-mercapto-beta,beta,-cyclopentamethylene propionyl, O-Et-Tyr2,Val4,Arg8]-vasopressin generated similar IC50 values suggesting that both these responses are mediated through the activation of a single V1a receptor subtype. 4. The onset of vasopressin-stimulated inositol-1,4,5-trisphosphate (Ins(1,4,5)P3) mass formation preceded [3H]-PtdBuOH accumulation indicating that PtdCh hydrolysis was activated subsequent to PtdIns(4,5)P2 breakdown. 5. The protein kinase C (PKC) activator, tetradecanoylphorbol acetate (TPA) also stimulated [3H]-PtdBuOH accumulation. Preincubation with the PKC inhibitor Ro-31-8220 abolished both TPA- and vasopressin-stimulated [3H]-PtdBuOH, suggesting that the intermediate activation of protein kinase C is involved in the regulation of PLD by vasopressin. 6. Pretreatment of the A10 VSMC with Ro-31-8220 (100 microM) also potentiated vasopressin-stimulated Ins(1,4,5)P3 mass formation.Therefore stimulation of PKC may have opposing roles in the regulation of agonist activation of PLC and PLD.7. Preincubation of the cells with EGTA, verapamil, or the receptor-operated calcium channel antagonist, SK&F 96365, reduced vasopressin-stimulated [3H]-PtdBuOH accumulation by approximately 30%, suggesting that influx of calcium has a significant role to play in the regulation of vasopressinstimulated PLD activity.

MeSH Terms
Animals Arginine Vasopressin/pharmacology Glycerophospholipids Hydrolysis Indoles/pharmacology Inositol 1,4,5-Trisphosphate/metabolism Mice Muscle, Smooth, Vascular/cytology,drug effects,metabolism Phosphatidic Acids/metabolism Phosphatidylcholines/metabolism Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositols/metabolism Rats Tetradecanoylphorbol Acetate/pharmacology Vasopressins/pharmacology
Chemicals
Glycerophospholipids Indoles Phosphatidic Acids Phosphatidylcholines Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositols phosphatidylbutanol Vasopressins Arginine Vasopressin Inositol 1,4,5-Trisphosphate Tetradecanoylphorbol Acetate Ro 31-8220
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Plevin R
Department of Biochemistry, University of Glasgow.
Stewart A
Paul A
Wakelam M J
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Article Info
Journal
British journal of pharmacology
Abbr.
Br J Pharmacol
ISSN
0007-1188
DOI
10.1111/j.1476-5381.1992.tb14471.x
Published
1992-09-00
Pages
109-15
Language
English
Region
England
NLM ID
7502536
PMCID
PMC1907616
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1111/j.1476-5381.1992.tb14471.x PMC Full Text View journal details
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